Additional remarks phenotype | Mutant/mutation
The mutant expresses a C-terminal 3xHA-tagged version of CDKrs
Protein (function)
CDKrs is a protein related to Cks1 and CksHs2, two CDK-associated proteins in Saccharomyces cerevisiae and and human, respectively. In the paper evidence is presented for an interaction between CRK5, SOC2 and CDKrs
From the Abstract: 'Cell cycle transitions are generally triggered by variation in the activity of cyclindependent kinases (CDKs) bound to cyclins. Malaria-causing parasites have a life cycle with unique cell-division cycles, and a repertoire of divergent CDKs and cyclins of poorly understood function and interdependency. We show that Plasmodium berghei CDK-related kinase 5 (CRK5), is a critical regulator of atypical mitosis in the gametogony and is required for mosquito transmission. It phosphorylates canonical CDK motifs of components in the pre-replicative complex and is essential for DNA replication. During a replicative cycle, CRK5 stably interacts with a single Plasmodium specific cyclin (SOC2), although we obtained no evidence of SOC2 cycling by transcription, translation or degradation. Our results provide evidence that during Plasmodium male gametogony, this divergent cyclin/CDK pair fills the functional space of other eukaryotic cell-cycle kinases controlling DNA replication.'
Phenotype
Not analysed in detail. CDKrs-3xHA in the nucleus of non-activated microgametocytes and throughout male gametogony, with a spindle-like localisation during mitoses, as found for CRK5. The mutant is used for mass-spec analyses of co-precipitated proteins in activated male gametocytes.
Additional information
Evidence in this study is presented that:
- CRK5 is a key regulator of gametogony and sporogony in the mosquito
- Phosphoproteome kinetics point to direct phosphorylation of the pre-replicative complex by CRK5
- CRK5 is required for both S- and M- phases during P. berghei gametogony
- CRK5 is part of an atypical nuclear cyclin/CDK complex
- CRK5, SOC2 ((PBANKA_1442200)and CDKrs (PBANKA_0824400; (CDK regulatory subunit, CDKrs) have a similar location and complementary functions during gametogony. CDKrs is a protein related to Cks1 and CksHs2, two CDK-associated proteins in Saccharomyces cerevisiae and and human, respectively.
- CRK5, SOC2 and CDKrs have a similar location and complementary functions during gametogony
- SOC2 expression does not follow a temporal cyclin pattern during gametogony and the CSC complex (CRK5/SOC2/CDKrs complex) is stable during the first round of mitosis
- CSC is dynamically phosphorylated during the first round of replication
- Live fluorescence microscopy identified CDKrs-GFP in the nucleus of non-activated microgametocytes and throughout male gametogony, with a spindle-like localisation during mitoses, as found for CRK5
- Ultrastructural analysis by electron microscopy of SOC2-KO and CDKrs-KO activated gametocytes confirms an early arrest during male gametogony as observed in CRK5-KO parasites.
Other mutants |